The Study Of Caffeic Acid Phenethyl Ester On The Prevention And Treatment Of Osteoporosis In Ovariectomized Mice

Background: Osteoporosis (osteoporosis, OP) is a systemic skeletaldisorder characterized by low bone mass, bone micro-structural damageleading to bone fragility, easy-to-fracture, occur in the elderly andmenopausal women. Current clinical treatment of osteoporosis drugs havea lot.Prevention and treatment of osteoporosis drugs are mainly is westernmedicine, while the herbal treatment of osteoporosis is currently in theemerging stage. Through our cooperative research laboratory and Australiafound that the active substance in propolis caffeic acid phenethyl ester(caffeic acid phenethyl ester, CAPE) include similar chemical structurewith NF-κB inhibitors . In vitro, the rankle pathway of CAPE inhibit thevalue-added and activities of osteoclast. By the early studies, we designthis animal experiments want to prove that CAPE could prevent and treatosteoporosis.Objective: To evaluate the prevention and treatment of CAPE on theosteoporosis of ovariectomized mice.Methods: 96 female 12-week C57BL/6J mice were randomly divided into16 groups (n = 6). Which, prevention of osteoporosis is divided into: 1week of sham group (SHAM) PBS group, simple group with ovariectomized (OVX)PBS group; 4 weeks and 7 weeks of SHAM-PBS group, OVX-PBS group, OVX withintraperitoneal injection of CAPE (0.1mg/ml), OVX with intraperitonealinjection of CAPE (0.2mg/ml). Treatment of osteoporosis were divided into:7 week of SHAM control group and OVX control group, the time of 7 weeksafter a successful modeling with intraperitoneal injection divided intofour groups: SHAM-PBS group, OVX-PBS group, OVX with intraperitonealinjection of CAPE (0.1mg / ml), OVX with intraperitoneal injection of CAPE(0.2mg/ml).Breeding the mice for 4 weeks, in the execution of the first3 days, 10 days with the mice injected calcein 0.5ml (1mg/mL). 96 micewere used 0.5% 0.3ml intraperitoneal injection of pentobarbitalanesthesia, after 5 minutes start heart blood, collected the bilateralfemur and tibia of mice. Detected by ELISA of serum estrogen, osteoblastmarkers ALP, RANKL. RT-PCR analysis the gene of right femur of RANKL, OPG,ALP expression levels. Observe the left tibia changes using microCT bone.observe the bilateral tibial changes by bone histomorphometry andstaining methods.Results: ELISA: 1, serum estrogen levels detected, removal of ovarianestrogen compared with SHAM, OVX group was significantly lower (p <0.05). 2, serum levels detected in the RANKLE, each group had no significantstatistical difference. 3, serum levels of ALP detected after 1 weeksignificantly decreased than ovariectomized group (p <0.05). 11-weektreatment group, the OVX group (0.1mg/ml) was significantly highercompared with SHAM group ALP (p <0.05).The RT-PCR results of mouse right femur showed: after 1 week the RANKLof OVX group was significantly higher (p <0.05). The 4 weeks of preventiongroup after the intraperitoneal injection of OVX group was significantlyhigher than that of ALP other two groups (p <0.05).The left tibia of MicroCT study found that : In the prevention andtreatment groups of the OVX group which injected intraperitoneally withCAPE (0.1mg/ml), the trabecular bone density, number, volume percentageincrease than OVX-PBS group (p <0.05), compared with the SHAM group wasnot significant.The research Using Bone histomorphometry and staining methodsanalysis bilateral tibial shows that: In the prevention group of the4-week group, CAPE (0.1mg/ml) in OVX osteoclast surface length of thepercentage of absorption, bone absorption coefficient than OVX-PBS groupincreased (p <0.05), compared with the SHAM group was not significant.Conclusion: The CAPE in vivo by inhibiting osteoclasts withvalue-added activities, play a role of prevention and treatment ofosteoporosis.